Identifying novel anti-infectives by high through-put screening in whole animals

通过对整体动物进行高通量筛选来鉴定新型抗感染药物

• 批准号: 7939581
• 负责人: Frederick M Ausubel
• 金额: $ 92.68万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-28 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7939581
• 关键词: Acinetobacter; Adhesions; Animals; Anti-Infective Agents; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Bacteria; Bypass; Caenorhabditis elegans; Chemicals; Disease; Drosophila genus; Drosophila melanogaster; Drug Delivery Systems; Enterobacter; Epithelial; Exhibits; Funding; Generations; Goals; Gram-Negative Bacteria; Health; Human; Human body; Immune; Incidence; Infection; Klebsiella; Knowledge; Life; Microbe; Microbial Biofilms; Modeling; Molecular; Molecular Target; Multi-Drug Resistance; Nematoda; Parasites; Pathway interactions; Pharmaceutical Preparations; Physiology; Poison; Process; Pseudomonas aeruginosa; Resistance development; Screening procedure; Signal Transduction; Speed; Testing; Tissues; Toxic effect; Virulence; Virus; Work; antimicrobial drug; drug discovery; efficacy testing; fungus; high throughput screening; immunoregulation; in vivo; innovation; killings; microbial; microorganism; mouse model; next generation; novel; pathogen; prevent; public health relevance; small molecule

DESCRIPTION (provided by applicant): Continuously emerging new and hard-to-treat microbes, and the growing incidence of multi-drug resistant infections pose formidable challenges to human health. Innovative approaches are urgently needed to speed up the discovery of new anti-infectives. Our aim is to achieve a paradigm shift in antimicrobial drug discovery by finding next generation anti-infectives that prevent disease by blocking pathogen adaptation to host physiology. To this end we propose using whole live animals for high throughput screening of small molecules. We have developed infection models in the nematode Caenorhabditis elegans that can be used to identify drugs that cure otherwise lethal infections. High throughput screening of nematodes in 384-well plates is followed by secondary screening in a more highly evolved model host, the fruit fly Drosophila melanogaster, increasing the likelihood of isolating drugs that will work in humans. Our approach is applicable to many different classes of microorganisms, including bacteria, viruses, fungi and parasites. It has several advantages over traditional drug discovery: (i) In addition to identifying conventional antibiotics, it will uncover entirely new classes of anti-infectives that only exhibit in vivo activity. Examples are "virulence blockers" and "immune escape blockers". (ii) Our approach is unbiased and requires no prior knowledge of potential drug targets or pathways. (iii) It bypasses the current bottleneck of toxicity/efficacy testing by automatically eliminating toxic compounds (because they would kill the nematodes), yielding quality hits with in vivo activity. (iv) It will identify compounds that prevent or mitigate microbial resistance development, or can be combined with antibiotic therapy, thereby increasing antibiotic efficacy. We predict that our approach can identify compounds that inhibit diverse aspects of virulence: (i) adhesion and colonization, (ii) epithelial barrier disruption, (iii) deep tissue invasion, (iv) biofilm formation, (v) avoidance of immune recognition, and (vi) modulation of immune signaling. Some of the molecular mechanisms underlying these processes are conserved across bacterial species. To establish proof-of-principle, we seek funding for discovering new anti-infectives against Pseudomonas aeruginosa, one of several gram-negative bacteria that have recently emerged in a multi-drug resistant form for which efficient antibiotics are either limited or not available. We plan to screen a large number of chemical compounds (250,000) to maximize the discovery of new classes of anti-infectives. Promising compounds will undergo characterization, efficacy testing in other gram-negative bacteria (Klebsiella, Acinetobacter, Enterobacter) and testing in mouse models of infection. For highly promising candidates we will attempt molecular target identification. PUBLIC HEALTH RELEVANCE: Microbes that cause disease are becoming resistant to antibiotics faster than we can find new ones, making many common infections untreatable and life threatening. The goal of our project is to find a way to identify a new generation of antibiotics. Rather than simply preventing bacteria from growing, these new sophisticated drugs will prevent disease by interfering with a microbe's ability to interact with the human body.

描述（申请人提供）：新的难治微生物不断出现，多重耐药感染发生率不断上升，给人类健康带来巨大挑战。迫切需要创新方法来加快新抗感染药物的发现。我们的目标是通过寻找下一代抗感染药物来实现抗菌药物发现的范式转变，这些抗感染药物通过阻止病原体对宿主生理的适应来预防疾病。为此，我们建议使用整个活体动物进行小分子的高通量筛选。我们开发了秀丽隐杆线虫感染模型，可用于识别治疗其他致命感染的药物。在 384 孔板中对线虫进行高通量筛选，然后在进化程度更高的模型宿主果蝇果蝇中进行二次筛选，从而增加分离对人类有效的药物的可能性。 我们的方法适用于许多不同类别的微生物，包括细菌、病毒、真菌和寄生虫。与传统药物发现相比，它具有以下几个优点：（i）除了识别传统抗生素之外，它还将发现仅表现出体内活性的全新类别的抗感染药物。例子是“毒力阻断剂”和“免疫逃逸阻断剂”。 (ii) 我们的方法是公正的，不需要事先了解潜在的药物靶点或途径。 (iii) 它通过自动消除有毒化合物（因为它们会杀死线虫），绕过了当前毒性/功效测试的瓶颈，产生具有体内活性的高质量命中。 (iv) 它将鉴定出能够预防或减轻微生物耐药性发展的化合物，或者可以与抗生素治疗相结合，从而提高抗生素疗效的化合物。 我们预测我们的方法可以识别抑制毒力各个方面的化合物：（i）粘附和定植，（ii）上皮屏障破坏，（iii）深层组织侵袭，（iv）生物膜形成，（v）避免免疫识别， (vi)免疫信号的调节。这些过程背后的一些分子机制在细菌物种之间是保守的。为了进行原理验证，我们寻求资金来发现针对铜绿假单胞菌的新型抗感染药物，铜绿假单胞菌是最近以多重耐药形式出现的几种革兰氏阴性细菌之一，有效的抗生素要么有限，要么无法获得。我们计划筛选大量化合物（250,000 种），以最大限度地发现新型抗感染药物。有前景的化合物将在其他革兰氏阴性菌（克雷伯氏菌、不动杆菌、肠杆菌）中进行表征、功效测试，并在小鼠感染模型中进行测试。对于非常有前途的候选者，我们将尝试分子靶点识别。 公共卫生相关性：引起疾病的微生物对抗生素产生耐药性的速度比我们发现新抗生素的速度更快，使得许多常见感染无法治疗并危及生命。我们项目的目标是找到一种方法来识别新一代抗生素。这些新型复杂药物不是简单地阻止细菌生长，而是通过干扰微生物与人体相互作用的能力来预防疾病。